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heat shock protein 27 regulates oxidative stress-induced apoptosis in cardiomyocytes:mechanisms via reactive oxygen species generation and Akt activation

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Chinese Medical Journal 2007年第24期120卷 2271-2277页

作者： LIU Li ZHANG Xiao-jin JIANG Su-rong DING Zheng-nian DING Guo-xian HUANG Jun CHENG Yun-lin Departments of Geriatrics First Affiliated Hospital of Nanjing Medical University Nanjing 210029 China Department of Anaesthesiology First Affiliated Hospital of Nanjing Medical University Nanjing 210029 China Department of Cardiology First Affiliated Hospital of Nanjing Medical University Nanjing 210029 China

Background Increased reactive oxygen species （ROS） formation, which in turn promotes cardiomyocytes apoptosis, is associated with the pathogenesis and progression of various cardiac diseases such as ischemia and heart failure. Recent studies have shown that over expression of heat shock protein 27 （Hsp27） confers resistance to cardiac ischemia/reperfusion injury. However, not much is known about the regulation of myocyte survival by Hsp27. Methods The rat cardiac cell line H9c2, with a stable overexpression of Hsp27, was established, with empty vector transfected H9c2 cells as controls. Following the cells challenged by Hydrogen Peroxide （H2O2）, lactate dehydrogenase （LDH） release, apoptosis, intracellular ROS, cell morphology, mitochondrial transmembrane potential and the activation of serine/threonine kinase Akt were determined. Results Along with marked suppression of H2O2-induced injury by Hsp27 overexpression in H9c2 cells, ROS generation and the loss of mitochondrial membrane potential were also significantly depressed. Furthermore, augmented Akt activation was observed in Hsp27 overexpressed H9c2 cells following H2O2 exposure. Conclusions Hsp27 inhibits oxidative stress-induced H9c2 damage and inhibition of ROS generation and the augmentation of Akt activation may be involved in the protective signaling.

关键词： heat shock protein 27 apoptosis superoxide mitochondrial potential Akt

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Paclitaxel-doxorubicin sequence is more effective in breast cancer cells with heat shock protein 27 overexpression

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Chinese Medical Journal 2008年第20期121卷 1975-1979页

作者： SHI Peng WANG Ming-ming JIANG Li-yu LIU Huan-tao SUN Jing-zhong Department of General Surgery Provincial Hospital Affiliated to Shandong University Jinan Shandong 250021 China Department of Breast Surgery Qilu Hospital Shandong University Jinan Shandong 250012 China

Background Cancer cells with overexpression of heat shock protein 27 （HSP27） are resistant to chemotherapeutic drug doxorubicin （Dox）. Paclitaxel （Pacl） was reported to suppress HSP27 expression in ovarian and uterine cancer cells. The purposes of this study were to investigate whether Pacl inhibits the expression of HSP27 in breast cancer cells, whether Pacl can sensitize breast cancer cells with HSP27 overexpression to Dox, and to define a more effective schedule for the combination of Dox with Pacl. Methods The HSP27 high-expressing human breast cancer cell lines, MCF-7 and MDA-MB-435, and the HSP27 low-expressing cell line, MDA-MB-231, were used in this study. The level of HSP27, topoisomerase （Topo） IIα and β expression were assessed by Western blotting. The cytotoxic activities of Dox, Pacl and combination of these two drugs were evaluated by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay and flow cytometric assays. Results Pacl （0.1 μmol/L） inhibited HSP27 expression by approximately 2-fold in MCF-7 and MDA-MB-435 cells, while up-regulating the level of topo IIα and β. In contrast, expression of HSP27 in MDA-MB-231 did not change significantly following Pacl treatment. There were synergistic effects in both treatment sequences （Pacl-Dox and Dox-Pacl） when Pacl was combined with Dox. Compared with those treated with the Dox-Pacl sequence, the Pacl-Dox sequence had a stronger effect in cancer cells with HSP27 overexpression, as MCF-7 and MDA-MB-435 treated with the Pacl-Dox sequence had lower viabilities and a higher apoptotic rate. Conclusions Paclitaxel significantly decreases the level of HSP27 in breast cancer cells overexpressing HSP27. In combination therapies, the Pacl-Dox sequence is more effective in clearing breast cancer cells with high HSP27 expression compared with the Dox-Pacl sequence.

关键词： apoptosis doxorubicin heat shock protein 27 paclitaxel drug resistance

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Expression of heat shock protein 27 in the esophageal tissue of rats with reflux esophagitis

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Chinese Medical Journal 2011年第15期124卷 2347-2353页

作者： ZHENG Chao-xu WANG Zhuo-qing LIN Wei-bin CHU Zhong-hua CHEN Liu-hua JI Zhuang-qi Department of General Surgery the First Affiliated Hospital Sun Yat-sen University Guangzhou Guangdong 510080 China Scientific Research Section the First Affiliated Hospital Sun Yat-sen University Guangzhou Guangdong 510080 China Department of General Surgery Hexian Memorial Hospital Panyu District Guangzhou Guangdong 511400 China Department of Gastrointestinal Surgery the Second Affiliated Hospital Sun Yat-sen University Gnangzhou Guangdong 510120 China

Background Little attention has been paid to the expression of heat shock protein 27 （HSP27） in patients with reflux esophagitis （RE）, and few studies of the importance of HSP27 in esophagitis have been carried out in animal models. This study aimed to explore the expression of HSP27 in the esophageal tissue of rats with RE. Methods Eighty female Wistar rats were randomly divided into experimental groups A and B and control groups C and D （n=20 in each group）. To establish RE, rats in the two experimental groups received pylorus and forestomach ligations, while rats in the control group received gastrostomy and gastric perforation repair. The rats in groups A and C were sacrificed 7 days after surgery, and the rats in groups B and D were sacrificed 14 days after surgery. In groups A and B, 10 and 8 rats were diagnosed with RE by pathological examination, respectively （they were included in groups A＇ and B＇, respectively）. The histopathological diagnosis of all the lower esophageal tissues in groups C and D was normal and 20 normal specimens were randomly selected for groups C＇ and D＇ with 10 specimens in each group. Macroscopic and microscopic esophagitis scores were assessed for the specimens in groups A＇ and B＇. Lower esophageal tissues were collected from groups A＇, B＇, C＇, and D＇, and paraffin-embedded slices were made using part of the tissues. The expression of HSP27 in the tissues was detected using the two-step streptavidin-peroxidase immunohistochemical method. Some collected tissues were frozen, and expressions of HSP27 mRNA were detected using fluorescence quantitative polymerase chain reaction （FQ-PCR）. Results Median macroscopic and microscopic esophagitis scores in groups A＇（n=10） and B＇（n=8） were 1.0 and 1.5, and 2.0 and 2.5, respectively. There were no significant differences in the macroscopic or microscopic esophagitis scores between the two groups （Z=-0.330, P=-0.741; Z=-0.142, P=-0.887, respectively）. Immunohistochemical staining showed that HSP27 was expressed in all layers of the esophagea

关键词： reflux esophagitis heat shock protein 27 rats

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Molecular cloning of human heat shock protein 27 and study of its protective effects on oxidative damage in rat cardiomyocte H9c2

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Journal of Nanjing Medical University 2005年第4期19卷 187-190页

作者： Li Liu Xiaojin Zhang Surong Jiang Xiang Gao Guoxain Ding Yunlin Cheng Department of Gerontology the First Affiliated Hospital of Nanjing Medical University Nanjing 210029 China Center of Model Animal Nanjing University Nanjing 210045China.

Objective： To clone human cardiac heat shock protein 27 （HSP27） gene and to determine the effects of HSP27 on the oxidative stress in rat cardiomyocyte cell line H9c2. Methods： Full length of HSP27 cDNA which got by RT-PCR was constructed into pCDNA3.1^＋ . The recombinant was transfected into rat cardiomyocyte cell line H9c2 and the stable trahsfection cell line was selected by G418. Then we observe the effects of HSP27 over-expression on LDH release and apoptosis induced H2O2 in H9c2. Results： ①pCDNA3.1^＋/HSP27 provided a sound expression of HSP27 in both 293T and H9c2. ②LDH releasing induced by 0, 100,250,500, 1000 μmol/L H2O2 in HSP27 over-expression group and wild type group were 0.396±0.017 vs. 0.390±0.01）9 （p 〉0.05）, 0.437±0. 014 vs. 0.416±0.015 （P〈0.05）, 0.471±0.018 vs. 0.417±0.009 （P 〈0.001）, 0.505±0.030 vs. 0.657± 0.022（P 〈0.001）, 0.547 ±0.027 and 0.661 ± 0.011（ P 〈 0. 001 ）, respectively. ③Apoptosis induced by 150 μmol/L H2O2 in HSP27 over-expression group and wild type group were （10.693± 1.122）% vs. （4.027 ± 1.628）%（ P 〈0.01）. Conclusion： We cloned and constructed human cardiac HSP27 gene successfully, and over-expression of human HSP27 could inhibit oxidative damage significantly in H9c2.

关键词： heat shock protein 27 Molecular Cloning Transfection Oxidafive Stress Apoptosis

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Comparison of Hsps Expression after Radio-frequency Field Exposure in Three Human Glioma Cell Lines

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Biomedical and Environmental Sciences 2009年第5期22卷 374-380页

作者： GUI -RONG DING, XIAO-WU WANG, KANG-CHU LI, LIAN-BO QIU, SHENG-LONG XU, JUAN TAN, AND GUO-ZHEN GUO1 Department of Radiation Medicine, Faculty of Preventive Medicine, Fourth Military Medical University, Xi’an 710032, Shaanxi, China Department of Radiation Medicine Faculty of Preventive Medicine Fourth Military Medical University Xi'an 710032 Shaanxi China

Objective To investigate and compare the effect of radio-frequency （RF） field exposure on expression of heat shock proteins （Hsps） in three human glioma cell lines （MO54, A172, and T98）. Methods Cells were exposed to sham or 1950 MHz continuous-wave for 1 h. Specific absorption rates （SARs） were 1 and 10 W/kg. Localization and expression of Hsp27 and phosphorylated Hsp27 （（78） Ser）（p-Hsp27） were examined by immunocytochemistry. Expression levels of Hsp27, p-Hs27, and Hsp70 were determined by Western blotting. Results The Hsp27 was primarily located within the cytoplasm, p-Hsp27 in both cytoplasm and nuclei of MO54, A172, and T98 cells. RF field exposure did not affect the distribution or expression of Hsp27. In addition, Western blotting showed no significant differences in protein expression of Hsp27 or HspT0 between sham- and RF field-exposed cells at a SAR of 1 W/kg and 10 W/kg for 1 h in three cells lines. Exposure to RF field at a SAR of 10 W/kg for 1 h slightly decreased the protein level of phosphorylated Hsp27 in MO54 cells. Conclusion The 1950 MHz RF field has only little or no apparent effect on Hsp70 and Hsp27 expression in MO54, A172, and T98 cells.

关键词： Radio-frequency field heat shock protein 70 heat shock protein 27 protein expression Phosphorylated Hsp27 Glioma cells

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Preconditioning crush increases the survival rate of motor neurons after spinal root avulsion

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Neural Regeneration Research 2014年第5期9卷 540-548页

作者： Lin Li Yizhi Zuo Jianwen He Department of Human Anatomy Nanjing Medical University

In a previous study, heat shock protein 27 was persistently upregulated in ventral motor neurons following nerve root avulsion or crush. Here, we examined whether the upregulation of heat shock protein 27 would increase the survival rate of motor neurons. Rats were divided into two groups： an avulsion-only group （avtflsion of the L4 lumbar nerve root only） and a crush-avulsion group （the L4 lumbar nerve root was crushed 1 week prior to the avulsion）. Immunofluores- cent staining revealed that the survival rate of motor neurons was significantly greater in the crush-avulsion group than in the avulsion-only group, and this difference remained for at least 5 weeks after avulsion. The higher neuronal survival rate may be explained by the upregulation of heat shock protein 27 expression in motor neurons in the crush-avulsion group. Further- more, preconditioning crush greatly attenuated the expression of nitric oxide synthase in the motor neurons. Our findings indicate that the neuroprotective action of preconditioning crush is mediated through the upregulation of heat shock protein 27 expression and the attenuation of neuronal nitric oxide synthase upregulation following avulsion.

关键词： nerve regeneration nerve root avulsion spinal nerve root heat shock protein 27 nitric oxide synthase motor neurons fluorescent antibody technique choline acetyltransferase a grant from Education Ministry of Jiangsu Province Excellent Discipline of Jiangsu Province neural regeneration

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Effects of Sodium Salicylate on the Expression of HSP27 protein during Oxidative Stress in Tissue-cultured Human Lens Epithelial Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2006年第6期26卷 753-755页

作者：王智周莉 Department of Ophthalmology Union Hospital Tongji Medical College Huazhong University of Science and Technology

The effects of sodium salicylate on the expression of heat shock protein 27 （HSP27） during oxidative stress in tissue-cultured human lens epithelial cells were investigated. Cultured human lens epithelial cells （HLB-3） were divided into 3 groups： control group （group A）, oxidation injury group （group B） and sodium salicylate group （group C）. Apoptosis of human lens epithelial cells cultured in vitro was induced in the presence of 150 μmol/L H2O2. Cells viability and the expression of HSP27 were analyzed. Viability of the cells was measured by methyl thiazole tetrazolium （MTT） chromatometry. The expression of HSP27 in HLB-3 cells was detected by using immunohistochemistry and image analysis system, Sodium salicylate could induce the expression of HSP27, and the cells viability in group C was significantly higher than in group B （0.2667±0.01414 vs 0.2150±0.01080, P=0.012〈0.05）. The average gray value of HSP27 in group B was less than that in group C （P=0.000〈0.05）. The increased expression of HSP27 by sodium salicylate might play an important role in the protection of hydrogen peroxide-induced injury of human lens epithelial cells, suggesting that sodium salicylate could suppress, at least in part, the apoptosis of human lens epithelial cells.

关键词： apoptosis human lens epithelial cells heat shock protein 27 sodium salicylate hydrogen peroxide

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微小RNA17-5p通过靶基因E2F1激活热休克蛋白27促进肝癌细胞转移(英文)

微小RNA17-5p通过靶基因E2F1激活热休克蛋白27促进肝癌细胞转移(...

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中国的遗传学研究——遗传学进步推动中国西部经济与社会发...

作者：王芳杨富张玲徐丹霍希松毕海珊孙树汉第二军医大学

miR-17-5p is overexpressed in hepatocellular carcinoma(HCC),but the specific regulatory mechanisms of miR-17-5p in HCC remain unknown.We investigated the molecular basis of miR-17-5p as an oncogene in human HCC cell lines.Our in vivo and in vitro data indicate that miR-17-5p up-regulates the migration and proliferation of HCC cells.Interestingly,proteomic and western blotting assays revealed that miR-17-5p significantly activates the p38 mitogen-activated protein kinase MAPK pathway and increases the phosphorylation of heat shock protein 27(HSP27).Our results also suggest that E2F1-dependent down-regulation of Wip1 regulates miR-17-5p-p38-HSP27 signaling.Furthermore,suppression of HSP27 expression by small interfering RNA or the p38 MAPK pathwayspecific inhibitor SB203580 decreases the migration of HCC cells overexpressing miR-17-5p but does not reduce their proliferation.Finally, we show that miR-17-5p expression correlates well with HSP27 status in primary human HCC tissues with metastasis.Conclusion:Our findings suggest that the p38 MAPK pathway plays a crucial role in miR-17-5p-induced phosphorylation of HSP27 and,as a consequence, phosphorylated HSP27 enhances the migration of HCC cells.Our data highlight an important role of miR-17-5p in the proliferation and migration of HCC cells and support the potential application of miR-17-5p in HCC therapy.

关键词： hepatocellular carcinoma miR-17-5p heat shock protein 27 p38 mitogen-activated protein kinase

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